Pub Date : 2026-01-01Epub Date: 2025-12-03DOI: 10.1016/j.jplph.2025.154671
Chi Zhang , Song-Qi Li , Pengwei Jing , Run-Xin Wu , Yu-Qing Ma , Ji-Xiao Wu , Ru-Feng Song , Wen-Cheng Liu
In ever-changing natural environments, plants have evolved precise and intricate regulatory networks to combat energy deprivation. Under limited energy supply, plants use autophagy to recycle cellular components and sustain vital processes. Autophagy represents an evolutionarily conserved mechanism operating at the subcellular level in eukaryotes. Reactive oxygen species (ROS), traditionally viewed as metabolic byproducts, exert concentration-dependent effects in plants: lower ROS in a controllable concentration range serve as signaling molecules modulating various aspects of plant growth, development and stress responses, whereas over-accumulating ROS induce oxidative damages, threatening plant growth and survival. Although the classification, metabolic dynamics, and multifaceted roles of ROS in plants have been extensively studied, the reciprocal regulatory interplay between ROS signaling and autophagy remains inadequately explored, particularly in plants. This review summarizes recent progress of plant ROS, autophagy, and their interplay, and also provides predictions and perspectives on the potential regulatory mechanisms between ROS and autophagy.
{"title":"Autophagy, ROS, and their interplay in plant adaptive responses","authors":"Chi Zhang , Song-Qi Li , Pengwei Jing , Run-Xin Wu , Yu-Qing Ma , Ji-Xiao Wu , Ru-Feng Song , Wen-Cheng Liu","doi":"10.1016/j.jplph.2025.154671","DOIUrl":"10.1016/j.jplph.2025.154671","url":null,"abstract":"<div><div>In ever-changing natural environments, plants have evolved precise and intricate regulatory networks to combat energy deprivation. Under limited energy supply, plants use autophagy to recycle cellular components and sustain vital processes. Autophagy represents an evolutionarily conserved mechanism operating at the subcellular level in eukaryotes. Reactive oxygen species (ROS), traditionally viewed as metabolic byproducts, exert concentration-dependent effects in plants: lower ROS in a controllable concentration range serve as signaling molecules modulating various aspects of plant growth, development and stress responses, whereas over-accumulating ROS induce oxidative damages, threatening plant growth and survival. Although the classification, metabolic dynamics, and multifaceted roles of ROS in plants have been extensively studied, the reciprocal regulatory interplay between ROS signaling and autophagy remains inadequately explored, particularly in plants. This review summarizes recent progress of plant ROS, autophagy, and their interplay, and also provides predictions and perspectives on the potential regulatory mechanisms between ROS and autophagy.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"316 ","pages":"Article 154671"},"PeriodicalIF":4.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145682371","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-01Epub Date: 2025-12-02DOI: 10.1016/j.jplph.2025.154668
Uwe Sonnewald , Quan-Sheng Qiu , Herbert J. Kronzucker
{"title":"Editorial: Squaring the circle: Challenges and breakthroughs in plant sciences","authors":"Uwe Sonnewald , Quan-Sheng Qiu , Herbert J. Kronzucker","doi":"10.1016/j.jplph.2025.154668","DOIUrl":"10.1016/j.jplph.2025.154668","url":null,"abstract":"","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"316 ","pages":"Article 154668"},"PeriodicalIF":4.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145682372","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-01-01Epub Date: 2025-12-09DOI: 10.1016/j.jplph.2025.154673
Ruiyuan Liu , Yan Zhan , Aiming Cui , Ying Qu , Wenjie Jin , Yan Du , Lixia Yu , Libin Zhou
Foxtail millet (Setaria italica (L.) Beauv) is an important crop for both basic research and sustainable dryland agriculture, due to its rapid growth, high seed yield, strong stress tolerance, and rich nutritional qualities. Multiple mutagenesis approaches have been used to broaden foxtail millet germplasm resources, among which heavy ion beam (HIB) irradiation has emerged as a powerful tool for inducing genetic variations in plant breeding. However, compared with other model species, genetic resources in foxtail millet remain limited, and the specific effects of HIB radiation on this crop are not yet fully understood. In this study, seeds of foxtail millet (Yugu 24) were irradiated using six different carbon-ion beams (CIB, 100 Gy–500 Gy), a commonly used HIB mutagen. Then, the mutagenic effects were evaluated in the M1 generation, conducted large-scale phenotype screening in the M2 generation, and analyzed the anatomical, physiological, and molecular mechanisms of stable leaf mutants. M1 plants exhibited dose-dependent responses, with the optimum CIB dose for Yugu 24 ranging from 110 Gy to 140 Gy. From 3100 M2 plants, we identified 56 individuals exhibiting obvious phenotypic variations, resulting in an overall mutation frequency of 1.81 % under CIB irradiation. High frequencies of leaf morphological mutations was observed in the M2 population. Stable leaf mutants were identified and further characterized in the M4 generation, which displayed distinct phenotypic variations, including changes in chloroplast structure, stomatal characteristics, and photosynthetic pigment content. Collectively, these findings establish a theoretical foundation for applying CIB irradiation in foxtail millet mutation breeding. Furthermore, the CIB-induced mutant library of Yugu 24provides a valuable resource for future functional genomics research on foxtail millet.
{"title":"Dose-dependent mutagenic effects of carbon-ion beams in foxtail millet: from phenotypic screening to physiological and molecular mechanisms","authors":"Ruiyuan Liu , Yan Zhan , Aiming Cui , Ying Qu , Wenjie Jin , Yan Du , Lixia Yu , Libin Zhou","doi":"10.1016/j.jplph.2025.154673","DOIUrl":"10.1016/j.jplph.2025.154673","url":null,"abstract":"<div><div>Foxtail millet (<em>Setaria italica</em> (L.) Beauv) is an important crop for both basic research and sustainable dryland agriculture, due to its rapid growth, high seed yield, strong stress tolerance, and rich nutritional qualities. Multiple mutagenesis approaches have been used to broaden foxtail millet germplasm resources, among which heavy ion beam (HIB) irradiation has emerged as a powerful tool for inducing genetic variations in plant breeding. However, compared with other model species, genetic resources in foxtail millet remain limited, and the specific effects of HIB radiation on this crop are not yet fully understood. In this study, seeds of foxtail millet (<em>Yugu 24</em>) were irradiated using six different carbon-ion beams (CIB, 100 Gy–500 Gy), a commonly used HIB mutagen. Then, the mutagenic effects were evaluated in the M<sub>1</sub> generation, conducted large-scale phenotype screening in the M<sub>2</sub> generation, and analyzed the anatomical, physiological, and molecular mechanisms of stable leaf mutants. M<sub>1</sub> plants exhibited dose-dependent responses, with the optimum CIB dose for <em>Yugu 24</em> ranging from 110 Gy to 140 Gy. From 3100 M2 plants, we identified 56 individuals exhibiting obvious phenotypic variations, resulting in an overall mutation frequency of 1.81 % under CIB irradiation. High frequencies of leaf morphological mutations was observed in the M<sub>2</sub> population. Stable leaf mutants were identified and further characterized in the M<sub>4</sub> generation, which displayed distinct phenotypic variations, including changes in chloroplast structure, stomatal characteristics, and photosynthetic pigment content. Collectively, these findings establish a theoretical foundation for applying CIB irradiation in foxtail millet mutation breeding. Furthermore, the CIB-induced mutant library of <em>Yugu 24</em>provides a valuable resource for future functional genomics research on foxtail millet.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"316 ","pages":"Article 154673"},"PeriodicalIF":4.1,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733584","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-11-01DOI: 10.1016/j.jplph.2025.154649
Kangjun Fan , Yue Wu , Yonghua Qin , Hongzao He , Lu Lv , Gang Li , Jiao Liu , Rui Qin , Hong Liu
SHORT INTERNODES (SHI)-related sequence (SRS) proteins are plant-specific transcription factors that modulate hormone biosynthesis and signalling. Their contribution to legume–rhizobium symbiosis, however, remains largely unexplored. Phylogenetic and collinearity analyses of legume SRS genes classified 12 subclasses and revealed soybean's evolutionary relationships, including large-scale gene duplication. GmSRS14 was specifically highly expressed in root nodules and localised in the nucleus only. Exogenous IAA modulates its expression at low concentrations (1 μM), while high concentrations (100 μM) decrease nodule expression. All ABA concentrations tested (10, 20 and 50 μM) inhibited nodule growth, nitrogenase activity and GmSRS14 expression. Functional validation via hairy root transformation demonstrated GmSRS14 overexpression (GmSRS14-OE) increased nodule number, weight, and nitrogenase activity, while GmSRS14 silencing (GmSRS14-RNAi) suppressed nodulation. This study provides a new idea for breeding soybean varieties with high efficiency of nitrogen fixation.
{"title":"Functional characterization of GmSRS14 in regulating root nodule development of soybean","authors":"Kangjun Fan , Yue Wu , Yonghua Qin , Hongzao He , Lu Lv , Gang Li , Jiao Liu , Rui Qin , Hong Liu","doi":"10.1016/j.jplph.2025.154649","DOIUrl":"10.1016/j.jplph.2025.154649","url":null,"abstract":"<div><div>SHORT INTERNODES (SHI)-related sequence (SRS) proteins are plant-specific transcription factors that modulate hormone biosynthesis and signalling. Their contribution to legume–rhizobium symbiosis, however, remains largely unexplored. Phylogenetic and collinearity analyses of legume <em>SRS</em> genes classified 12 subclasses and revealed soybean's evolutionary relationships, including large-scale gene duplication. <em>GmSRS14</em> was specifically highly expressed in root nodules and localised in the nucleus only. Exogenous IAA modulates its expression at low concentrations (1 μM), while high concentrations (100 μM) decrease nodule expression. All ABA concentrations tested (10, 20 and 50 μM) inhibited nodule growth, nitrogenase activity and <em>GmSRS14</em> expression. Functional validation via hairy root transformation demonstrated <em>GmSRS14</em> overexpression (<em>GmSRS14</em>-OE) increased nodule number, weight, and nitrogenase activity, while <em>GmSRS14</em> silencing (<em>GmSRS14-</em>RNAi) suppressed nodulation. This study provides a new idea for breeding soybean varieties with high efficiency of nitrogen fixation.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"315 ","pages":"Article 154649"},"PeriodicalIF":4.1,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145459031","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cytokinins (CKs) exist in various forms within potato plants, among which the active CKs account for only a minute fraction but play crucial roles in tuber development. In the present study, in vitro potato stolons were exposed to graded concentrations of active CK N6-(Δ2-isopentenyl)-adenine (2ip) and CK-biosynthesis inhibitor lovastatin, and the phenotypic and endogenous phytohormone dynamics during tuber development were investigated. The results showed that low 2ip concentrations promoted tuberization, with 0.1 μM 2ip exhibiting the strongest inductive effect. The initial time of tuberization was advanced, and the tuberization rate, tuber fresh weight, and tuber diameter significantly increased. With increasing 2ip concentrations, higher levels (>50 μM) inhibited tuberization and markedly elevated the length-to-width ratio of tubers. Inhibiting CK biosynthesis by lovastatin also inhibited tuberization, and even induced the formation of abnormal tubers. Treatment with 0.1 μM 2ip shifted the endogenous hormone balance toward a state that favors tuber formation and development. Levels of active CKs (iP, tZ, and DHZ), IAA, and SA significantly increased, whereas the contents of total jasmonates (JA, JA-Ile, and cis-OPDA), ABA, and inactive CKs (iPR and cZR) decreased. The ratios of active CKs to GA3, ABA, or JA, as well as the IAA/ABA and IAA/GA3 ratios, significantly increased. Inhibition of CK biosynthesis elicited changes in CKs, JAs, and ABA levels, as well as in the associated phytohormone ratios, that were opposite to those observed with 0.1 μM 2ip treatment. Thus this study revealed the specific physiological roles of active CKs in tuber development and provided insights into the mechanisms of tuber development regulated by CKs.
{"title":"Cytokinin-induced phenotypic and endogenous phytohormonal dynamics during potato (Solanum tuberosum L.) tuber development in vitro","authors":"Lixiang Cheng, Jianlong Yuan, Lulu Xia, Zhensan Tang, Feng Zhang","doi":"10.1016/j.jplph.2025.154632","DOIUrl":"10.1016/j.jplph.2025.154632","url":null,"abstract":"<div><div>Cytokinins (CKs) exist in various forms within potato plants, among which the active CKs account for only a minute fraction but play crucial roles in tuber development. In the present study, <em>in vitro</em> potato stolons were exposed to graded concentrations of active CK N<sup>6</sup>-(Δ<sup>2</sup>-isopentenyl)-adenine (2ip) and CK-biosynthesis inhibitor lovastatin, and the phenotypic and endogenous phytohormone dynamics during tuber development were investigated. The results showed that low 2ip concentrations promoted tuberization, with 0.1 μM 2ip exhibiting the strongest inductive effect. The initial time of tuberization was advanced, and the tuberization rate, tuber fresh weight, and tuber diameter significantly increased. With increasing 2ip concentrations, higher levels (>50 μM) inhibited tuberization and markedly elevated the length-to-width ratio of tubers. Inhibiting CK biosynthesis by lovastatin also inhibited tuberization, and even induced the formation of abnormal tubers. Treatment with 0.1 μM 2ip shifted the endogenous hormone balance toward a state that favors tuber formation and development. Levels of active CKs (iP, tZ, and DHZ), IAA, and SA significantly increased, whereas the contents of total jasmonates (JA, JA-Ile, and <em>cis</em>-OPDA), ABA, and inactive CKs (iPR and cZR) decreased. The ratios of active CKs to GA<sub>3</sub>, ABA, or JA, as well as the IAA/ABA and IAA/GA<sub>3</sub> ratios, significantly increased. Inhibition of CK biosynthesis elicited changes in CKs, JAs, and ABA levels, as well as in the associated phytohormone ratios, that were opposite to those observed with 0.1 μM 2ip treatment. Thus this study revealed the specific physiological roles of active CKs in tuber development and provided insights into the mechanisms of tuber development regulated by CKs.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"315 ","pages":"Article 154632"},"PeriodicalIF":4.1,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145292518","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-10-29DOI: 10.1016/j.jplph.2025.154648
Songsong Jin , Xinling Zhong , Zhangli Hu , Zhonghao Jiang
Calcium (Ca2+) plays versatile roles in plant growth and development, as well as in responses to environmental stimuli. Abiotic stressors, including abnormal temperature, drought, salt, heavy metals, and flooding, induce instantaneous and rapid free cytosolic Ca2+ ([Ca2+]cyt) elevation, known as Ca2+ signatures. Ca2+ signatures are stress-specific and regulated by Ca2+ flux. Ca2+ flux contains Ca2+ influx, which initiates Ca2+ signatures, and Ca2+ efflux, which terminates them. Ca2+ flux is achieved through the co-operation of Ca2+ channels and pumps. Here, we highlight recent advances in Ca2+ flux and Ca2+ channel functions in plant responses to abiotic stress. Ca2+ influx channels in plants include the hyperosmolarity-gated calcium-permeable channel family of proteins (OSCAs), glutamate receptor-like channels (GLRs), cyclic-nucleotide-gated calcium channels (CNGCs), annexins (ANNs), two-pore channels (TPCs), Piezo channels (PZOs), Mid1-complement activity protein channels (MCAs), and mechanosensitive channels of small conductance (MscS)-like proteins (MSLs). Ca2+ efflux channels mainly contain Ca2+-ATPases and Ca2+ exchangers. Most Ca2+ channels have been found to participate in plant responses to single abiotic stress, whereas some are reported to be involved in responses to multiple abiotic stresses. This improved knowledge advances our understanding of Ca2+ signaling in plant responses to abiotic stress and offers new strategies for cultivating stress-resilient crops.
{"title":"Ca2+ flux in plant responses to abiotic stress","authors":"Songsong Jin , Xinling Zhong , Zhangli Hu , Zhonghao Jiang","doi":"10.1016/j.jplph.2025.154648","DOIUrl":"10.1016/j.jplph.2025.154648","url":null,"abstract":"<div><div>Calcium (Ca<sup>2+</sup>) plays versatile roles in plant growth and development, as well as in responses to environmental stimuli. Abiotic stressors, including abnormal temperature, drought, salt, heavy metals, and flooding, induce instantaneous and rapid free cytosolic Ca<sup>2+</sup> ([Ca<sup>2+</sup>]<sub>cyt</sub>) elevation, known as Ca<sup>2+</sup> signatures. Ca<sup>2+</sup> signatures are stress-specific and regulated by Ca<sup>2+</sup> flux. Ca<sup>2+</sup> flux contains Ca<sup>2+</sup> influx, which initiates Ca<sup>2+</sup> signatures, and Ca<sup>2+</sup> efflux, which terminates them. Ca<sup>2+</sup> flux is achieved through the co-operation of Ca<sup>2+</sup> channels and pumps. Here, we highlight recent advances in Ca<sup>2+</sup> flux and Ca<sup>2+</sup> channel functions in plant responses to abiotic stress. Ca<sup>2+</sup> influx channels in plants include the hyperosmolarity-gated calcium-permeable channel family of proteins (OSCAs), glutamate receptor-like channels (GLRs), cyclic-nucleotide-gated calcium channels (CNGCs), annexins (ANNs), two-pore channels (TPCs), Piezo channels (PZOs), Mid1-complement activity protein channels (MCAs), and mechanosensitive channels of small conductance (MscS)-like proteins (MSLs). Ca<sup>2+</sup> efflux channels mainly contain Ca<sup>2+</sup>-ATPases and Ca<sup>2+</sup> exchangers. Most Ca<sup>2+</sup> channels have been found to participate in plant responses to single abiotic stress, whereas some are reported to be involved in responses to multiple abiotic stresses. This improved knowledge advances our understanding of Ca<sup>2+</sup> signaling in plant responses to abiotic stress and offers new strategies for cultivating stress-resilient crops.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"315 ","pages":"Article 154648"},"PeriodicalIF":4.1,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145445179","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-10-21DOI: 10.1016/j.jplph.2025.154639
Mingyue Xu, Wenbin Su, Delight Hwarari, Yinyue Zuo, Zhaodong Hao, Jisen Shi, Jinhui Chen, Liming Yang
An efficient genetic transformation system is imperative for advancing gene functional studies and molecular breeding in horticultural tree species. Traditional Agrobacterium tumefaciens-mediated methods have encountered significant technical challenges when applied to tree species, particularly Liriodendron hybrids. The high costs, and extended transformation cycles associated with this method have substantially limited its utility in gene functional analysis and breeding applications. In response to these challenges, this study presents the development of a streamlined, rapid, and efficient Agrobacterium rhizogenes-mediated transformation system tailored specifically for Liriodendron hybrids, an important ornamental and timber tree species. Among the three Agrobacterium rhizogenes strains (K599, MSU440, and C58C1) evaluated in this study, K599 demonstrated the highest transformation efficiency, reaching 46.09 % for inducing hairy roots from apical bud incisions. Further optimization of the system revealed that 2 days and two-month-old seedlings were the most suitable co-culture duration and explants, yielding a peak transformation efficiency of 60.38 %, 53.12 %, respectively. The applicability of this transformation system was validated across various Liriodendron hybrids genotypes and Liriodendron tulipifera, with transformation efficiencies ranging from 15.47 % to 60.63 %. Additionally, the system was effectively employed for subcellular localization analysis, which confirmed that the aquaporin (AQP) protein. LhAQP1 is localized in the plasma membrane and exhibits enrichment in the vascular tissues of the hairy roots. Notably, this study marks the first application of the Agrobacterium rhizogenes-mediated system for CRISPR/Cas9-mediated gene editing in Liriodendron hybrids, successfully achieving targeted mutagenesis of the LhAQP1 gene and establishing the feasibility of gene editing within this species. In addition, the hairy root-based transformation system was employed to investigate the functional role of LhAQP1 in the improved Liriodendron variety ‘Nanlin-Jinsen E1’ by comparing wild-type, LhAQP1-overexpressing and gene-edited lines generated via the CRISPR/Cas9. LhAQP1-overexpression significantly promoted plant growth and drought tolerance, whereas pYLCRISPR-LhAQP1 increases dehydration sensitivity, underscoring the essential role of LhAQP1 in water stress adaptation. Optimized transformation platform represents a crucial advancement for functional genomics and molecular breeding efforts in woody plants.
{"title":"An efficient genetic transformation and gene editing system mediated by Agrobacterium rhizogenes for Liriodendron hybrid and its application","authors":"Mingyue Xu, Wenbin Su, Delight Hwarari, Yinyue Zuo, Zhaodong Hao, Jisen Shi, Jinhui Chen, Liming Yang","doi":"10.1016/j.jplph.2025.154639","DOIUrl":"10.1016/j.jplph.2025.154639","url":null,"abstract":"<div><div>An efficient genetic transformation system is imperative for advancing gene functional studies and molecular breeding in horticultural tree species. Traditional <em>Agrobacterium tumefaciens</em>-mediated methods have encountered significant technical challenges when applied to tree species, particularly <em>Liriodendron</em> hybrids. The high costs, and extended transformation cycles associated with this method have substantially limited its utility in gene functional analysis and breeding applications. In response to these challenges, this study presents the development of a streamlined, rapid, and efficient <em>Agrobacterium rhizogenes</em>-mediated transformation system tailored specifically for <em>Liriodendron</em> hybrids, an important ornamental and timber tree species. Among the three <em>Agrobacterium rhizogenes</em> strains (K599, MSU440, and C58C1) evaluated in this study, K599 demonstrated the highest transformation efficiency, reaching 46.09 % for inducing hairy roots from apical bud incisions. Further optimization of the system revealed that 2 days and two-month-old seedlings were the most suitable co-culture duration and explants, yielding a peak transformation efficiency of 60.38 %, 53.12 %, respectively. The applicability of this transformation system was validated across various <em>Liriodendron</em> hybrids genotypes and <em>Liriodendron tulipifera</em>, with transformation efficiencies ranging from 15.47 % to 60.63 %. Additionally, the system was effectively employed for subcellular localization analysis, which confirmed that the aquaporin (AQP) protein. LhAQP1 is localized in the plasma membrane and exhibits enrichment in the vascular tissues of the hairy roots. Notably, this study marks the first application of the <em>Agrobacterium rhizogenes</em>-mediated system for CRISPR/Cas9-mediated gene editing in <em>Liriodendron</em> hybrids, successfully achieving targeted mutagenesis of the <em>LhAQP1</em> gene and establishing the feasibility of gene editing within this species. In addition, the hairy root-based transformation system was employed to investigate the functional role of <em>LhAQP1</em> in the improved <em>Liriodendron</em> variety ‘Nanlin-Jinsen E1’ by comparing wild-type, <em>LhAQP1</em>-overexpressing and gene-edited lines generated via the CRISPR/Cas9. <em>LhAQP1</em>-overexpression significantly promoted plant growth and drought tolerance, whereas pYLCRISPR-LhAQP1 increases dehydration sensitivity, underscoring the essential role of <em>LhAQP1</em> in water stress adaptation. Optimized transformation platform represents a crucial advancement for functional genomics and molecular breeding efforts in woody plants.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"315 ","pages":"Article 154639"},"PeriodicalIF":4.1,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145377691","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-10-15DOI: 10.1016/j.jplph.2025.154635
Chunyi Ye , Weijia Kong , Yue Li , Huiyun Song , Ziyi Tian , Chunxia Lei , Pei Li
Background
Protoplasts are widely used in the fields of genetic transformation, physiology, and biochemistry, as they can easily absorb exogenous substances. The development and an efficient protoplast isolation and transient transformation system are essential for molecular biology and related research. Toona ciliata, valued for its high-quality and vividly colored wood, represents an economically significant species. In order to promote efficient breeding of the precious fast-growing tree, the establishment of a protoplast isolation and transient transformation system for T. ciliata is particularly important.
Results
The native protoplast isolation system established in this study used 0.3 g of leaves of two-month-old T. ciliata seedlings as the separation material, with an enzyme solution composed of 15 g/L Cellulase R-10 + 15 g/L Macerozyme R-10 + 0.6 M mannitol +10 mM MES +1 mM CaCl2 + 0.1 % BSA. Protoplasts were isolated in the dark at room temperature with gentle shaking (50 rpm) for 10 h, yielding (89.17 ± 7.21) × 106 protoplasts per gram of fresh weight with a viability of 92.62 ± 0.75 % (n = 3). For transient transformation, the optimal conditions included 40 % PEG, a plasmid concentration of 30 μg/μL, and a 30 min incubation in the dark, resulting in a transformation efficiency of 29.02 ± 6.13 % (n = 3). This highly efficient native protoplast-based transient expression system was successfully applied to determine the subcellular localization of 1-deoxy-D-xylulose 5-phosphate synthase (DXS), a rate-limiting enzyme in the terpenoid biosynthesis pathway in T. ciliata.
Conclusions
The established protoplast isolation and transient expression system provides a foundation for the subsequent identification of gene function and mechanism research, and provides a reliable research platform for the molecular breeding of T. ciliata, supporting future genetic improvement efforts.
背景:原生质体易吸收外源物质,在遗传转化、生理生化等领域有着广泛的应用。建立高效的原生质体分离和瞬时转化系统是分子生物学及相关研究的基础。香椿纤毛以其高质量和色彩鲜艳的木材而受到重视,是一种具有重要经济意义的物种。为了促进这一珍贵的速生树种的高效育种,建立一种原生质体分离和瞬时转化体系显得尤为重要。结果:本研究建立的原生质体分离体系以2月龄毛缕青苗叶片0.3 g为分离材料,酶液为15 g/L纤维素酶R-10 + 15 g/L宏观酶R-10 + 0.6 M甘露醇+10 mM MES +1 mM CaCl2 + 0.1% BSA。在室温暗摇(50 rpm)条件下分离原生质体10 h,每克鲜重产生(89.17±7.21)× 106个原生质体,活力为92.62±0.75% (n = 3)。瞬时转化的最佳条件为PEG含量为40%,质粒浓度为30 μg/μL,暗培养30 min,转化效率为29.02±6.13% (n = 3)。这种基于原生质体的高效瞬时表达系统成功地测定了纤毛t萜类生物合成途径中的限速酶1-脱氧-d -木lulose 5-磷酸合酶(DXS)的亚细胞定位。结论:建立的原生质体分离及瞬时表达体系为后续基因功能鉴定及机制研究奠定了基础,为纤毛虱分子育种提供了可靠的研究平台,为今后的遗传改良工作提供支持。
{"title":"Protoplast isolation and transient expression in the precious and economically important tree Toona ciliata","authors":"Chunyi Ye , Weijia Kong , Yue Li , Huiyun Song , Ziyi Tian , Chunxia Lei , Pei Li","doi":"10.1016/j.jplph.2025.154635","DOIUrl":"10.1016/j.jplph.2025.154635","url":null,"abstract":"<div><h3>Background</h3><div>Protoplasts are widely used in the fields of genetic transformation, physiology, and biochemistry, as they can easily absorb exogenous substances. The development and an efficient protoplast isolation and transient transformation system are essential for molecular biology and related research. <em>Toona ciliata</em>, valued for its high-quality and vividly colored wood, represents an economically significant species. In order to promote efficient breeding of the precious fast-growing tree, the establishment of a protoplast isolation and transient transformation system for <em>T. ciliata</em> is particularly important.</div></div><div><h3>Results</h3><div>The native protoplast isolation system established in this study used 0.3 g of leaves of two-month-old <em>T. ciliata</em> seedlings as the separation material, with an enzyme solution composed of 15 g/L Cellulase R-10 + 15 g/L Macerozyme R-10 + 0.6 M mannitol +10 mM MES +1 mM CaCl<sub>2</sub> + 0.1 % BSA. Protoplasts were isolated in the dark at room temperature with gentle shaking (50 rpm) for 10 h, yielding (89.17 ± 7.21) × 10<sup>6</sup> protoplasts per gram of fresh weight with a viability of 92.62 ± 0.75 % (n = 3). For transient transformation, the optimal conditions included 40 % PEG, a plasmid concentration of 30 μg/μL, and a 30 min incubation in the dark, resulting in a transformation efficiency of 29.02 ± 6.13 % (n = 3). This highly efficient native protoplast-based transient expression system was successfully applied to determine the subcellular localization of 1-deoxy-D-xylulose 5-phosphate synthase (DXS), a rate-limiting enzyme in the terpenoid biosynthesis pathway in <em>T. ciliata</em>.</div></div><div><h3>Conclusions</h3><div>The established protoplast isolation and transient expression system provides a foundation for the subsequent identification of gene function and mechanism research, and provides a reliable research platform for the molecular breeding of <em>T. ciliata</em>, supporting future genetic improvement efforts.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"315 ","pages":"Article 154635"},"PeriodicalIF":4.1,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145329605","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-10-25DOI: 10.1016/j.jplph.2025.154638
Bing Cui , Yiran Xu , Yancan Li , Jie Song , Faujiah Nurhasanah Ritonga , Jianwei Gao , Jingjuan Li
Soil salinization is one of the critical challenges facing global agriculture, seriously affecting crop growth and yields. As the primary organs for absorbing water and nutrients, plant roots play a pivotal role in responding to salt stress. In recent years, researchers have uncovered diverse functional mechanisms underlying root-mediated salt tolerance through multi-dimensional studies, such as gene regulation, physiological and biochemical mechanisms, and ecological adaptation. This article reviews the integrated functions of plant roots in salt tolerance from multiple perspectives, including root structure and function, physiological and biochemical responses of roots, root-shoot coordination, root perception and signal transduction of salt stress, rhizosphere microbial synergy, nutrient elements, and the application of phenotypic techniques. Such knowledge is expected to provide a theoretical foundation for breeding salt-tolerant crop varieties.
{"title":"The integrated function of roots in plant salt tolerance","authors":"Bing Cui , Yiran Xu , Yancan Li , Jie Song , Faujiah Nurhasanah Ritonga , Jianwei Gao , Jingjuan Li","doi":"10.1016/j.jplph.2025.154638","DOIUrl":"10.1016/j.jplph.2025.154638","url":null,"abstract":"<div><div>Soil salinization is one of the critical challenges facing global agriculture, seriously affecting crop growth and yields. As the primary organs for absorbing water and nutrients, plant roots play a pivotal role in responding to salt stress. In recent years, researchers have uncovered diverse functional mechanisms underlying root-mediated salt tolerance through multi-dimensional studies, such as gene regulation, physiological and biochemical mechanisms, and ecological adaptation. This article reviews the integrated functions of plant roots in salt tolerance from multiple perspectives, including root structure and function, physiological and biochemical responses of roots, root-shoot coordination, root perception and signal transduction of salt stress, rhizosphere microbial synergy, nutrient elements, and the application of phenotypic techniques. Such knowledge is expected to provide a theoretical foundation for breeding salt-tolerant crop varieties.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"315 ","pages":"Article 154638"},"PeriodicalIF":4.1,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145409239","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-12-01Epub Date: 2025-10-15DOI: 10.1016/j.jplph.2025.154636
Cassia Ayumi Takahashi , Rafael Silva Oliveira , Helenice Mercier
In tank-forming epiphytic bromeliads, two distinct growth stages can be easily identified, each characterized by specific adaptive traits for capturing nutrients such as inorganic or organic nitrogen sources: (a) the juvenile stage (atmospheric form), during which the bromeliad absorbs nutrients dissolved in rainwater through its leaves and roots; and (b) the adult stage (tank form), in which overlapping leaves form a reservoir that enables the accumulation of water and nutrients among the leaf bases. This study investigated differences in nitrogen absorption, translocation, and assimilation between these two growth stages of Vriesea gigantea. Atmospheric and tank-form bromeliads were supplied with solutions containing 15NO3−, 15NH4+, or [U-15N]urea. Leaves and roots were harvested at six different time points and used for enzymatic activity assays (urease, nitrate reductase, glutamine synthetase, glutamate dehydrogenase) and endogenous content quantifications (ammonium, nitrate, and 15N abundance). Ammonium and urea were the main nitrogen sources utilized by both growth forms. However, they were not absorbed and assimilated with equal efficiency: atmospheric bromeliads used ammonium more efficiently, whereas tank bromeliads utilized urea better. Although nitrate was the least absorbed source in both plants, atmospheric bromeliads showed faster uptake and assimilation. These findings suggest that inorganic nitrogen sources may be more readily available to epiphytic bromeliads during their juvenile phase, which could explain why they are physiologically better adapted to absorb and metabolize them. In the adult stage, organic nitrogen sources may become more accessible to V. gigantea, as the tank structure facilitates the accumulation of decomposing organic matter.
{"title":"Dynamics of nitrogen absorption, translocation, and assimilation depend on the growth stages of tank-forming epiphytic bromeliads","authors":"Cassia Ayumi Takahashi , Rafael Silva Oliveira , Helenice Mercier","doi":"10.1016/j.jplph.2025.154636","DOIUrl":"10.1016/j.jplph.2025.154636","url":null,"abstract":"<div><div>In tank-forming epiphytic bromeliads, two distinct growth stages can be easily identified, each characterized by specific adaptive traits for capturing nutrients such as inorganic or organic nitrogen sources: (a) the juvenile stage (atmospheric form), during which the bromeliad absorbs nutrients dissolved in rainwater through its leaves and roots; and (b) the adult stage (tank form), in which overlapping leaves form a reservoir that enables the accumulation of water and nutrients among the leaf bases. This study investigated differences in nitrogen absorption, translocation, and assimilation between these two growth stages of <em>Vriesea gigantea.</em> Atmospheric and tank-form bromeliads were supplied with solutions containing <sup>15</sup>NO<sub>3</sub><sup>−</sup>, <sup>15</sup>NH<sub>4</sub><sup>+</sup>, or [U-<sup>15</sup>N]urea. Leaves and roots were harvested at six different time points and used for enzymatic activity assays (urease, nitrate reductase, glutamine synthetase, glutamate dehydrogenase) and endogenous content quantifications (ammonium, nitrate, and <sup>15</sup>N abundance). Ammonium and urea were the main nitrogen sources utilized by both growth forms. However, they were not absorbed and assimilated with equal efficiency: atmospheric bromeliads used ammonium more efficiently, whereas tank bromeliads utilized urea better. Although nitrate was the least absorbed source in both plants, atmospheric bromeliads showed faster uptake and assimilation. These findings suggest that inorganic nitrogen sources may be more readily available to epiphytic bromeliads during their juvenile phase, which could explain why they are physiologically better adapted to absorb and metabolize them. In the adult stage, organic nitrogen sources may become more accessible to <em>V. gigantea</em>, as the tank structure facilitates the accumulation of decomposing organic matter.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"315 ","pages":"Article 154636"},"PeriodicalIF":4.1,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145401311","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}