Journal of plant physiology最新文献

{"title":"MbWRKY50 confers cold and drought tolerance through upregulating antioxidant capacity associated with ROS scavenging","authors":"Xinhui Wang ,&nbsp;Yingnan Li ,&nbsp;Zhuo Chen ,&nbsp;Longfeng Li ,&nbsp;Qiqi Li ,&nbsp;Zihan Geng ,&nbsp;Wanda Liu ,&nbsp;Ruining Hou ,&nbsp;Lihua Zhang ,&nbsp;Deguo Han","doi":"10.1016/j.jplph.2025.154526","DOIUrl":"10.1016/j.jplph.2025.154526","url":null,"abstract":"<div><div>Plant growth and development can be impacted by abiotic factors, including low temperature and dryness. Numerous studies have shown that plant responses to stress are largely influenced by the WRKY transcription factors (TFs). However, there are few studies on the role of WRKY genes in the stress response of <em>Malus</em> plants. In this experiment, <em>Malus baccata</em> (L.) Borkh was selected as the material, and the WRKY family gene <em>MbWRKY50</em> was cloned using a gene cloning technique. Phylogenetic tree analysis revealed that MbWRKY50 and MdWRKY50 have the highest homology. Furthermore, a green fluorescent protein (GFP) fusion protein expression assay revealed that the MbWRKY50 protein is located in the nucleus. The results of RT-qPCR showed that the expression of <em>MbWRKY50</em> was increased in the roots and fully grown leaves of <em>M. baccata</em>, and the response to low temperature and drought environment was enhanced<em>.</em> The <em>MbWRKY50</em> gene was transferred into tomato, which could better adapt to the cold and dry living conditions. In contrast to wild-type (WT) and untransformed (UL) tomato lines, overexpression of <em>MbWRKY50</em> boosts the activities of superoxide dismutase (SOD) and peroxidase (POD). Moreover, it leads to a notable reduction in the concentrations of malondialdehyde (MDA), hydrogen peroxide (H₂O₂), and superoxide anion free radicals (O₂⁻). The results showed that MbWRKY50 activated the expression levels of <em>LeABI3</em>, <em>LeNCED1</em>, <em>LeABF4</em>, <em>LeDREB1</em>, <em>LeCBF1</em>, and <em>LeCBF3</em> by binding to cold binding factor/dehydration response element (CBF/DREB) or participating in ABA synthesis, thereby enhancing the resistance of transgenic tomatoes to low temperature and drought stress.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"310 ","pages":"Article 154526"},"PeriodicalIF":4.0,"publicationDate":"2025-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144147918","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}

{"title":"Melatonin improves aluminum tolerance in soybean through modification of cell wall and vacuolar compartmentalization of aluminum","authors":"Huanan Li ,&nbsp;Ziyu Jia ,&nbsp;Kunxia Cheng ,&nbsp;Luyu Wang ,&nbsp;Junjun Huang ,&nbsp;Huahua Wang","doi":"10.1016/j.jplph.2025.154525","DOIUrl":"10.1016/j.jplph.2025.154525","url":null,"abstract":"<div><div>Aluminum (Al) toxicity has a significant adverse impact on plant growth and crop yield. Melatonin (MT) is involved in plant responses to various environmental stresses. However, the role of MT in mitigating Al toxicity remains largely unknown in soybean (<em>Glycine</em> max L.). In this study, the findings indicate that MT application alleviates Al-induced root growth inhibition and reduces Al accumulation in the cell wall. MT application under Al stress decreased the pectin and hemicellulose 1 content in the root tip cell wall and increased pectin methyl esterification, leading to reduced Al binding in the cell wall fractions. Additionally, MT treatment under Al stress inhibited lignin synthesis in the root tip cell wall, thereby alleviating the increased rigidity of cell wall and promoting its expansion. Furthermore, MT application under Al stress modulated the expression of Al transport-related genes (<em>GmCDT3</em>, <em>GmNrat1</em>, <em>GmIREG3</em> and <em>GmALS1</em>), reducing cytoplasmic Al accumulation and enhancing vacuolar Al sequestration. Taken together, these findings suggest that MT mitigates Al toxicity in soybean by reducing Al deposition in the cell wall and enhancing Al sequestration in the vacuole. This study offers clues for enhancing crop resistance to Al toxicity in acidic soils.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"310 ","pages":"Article 154525"},"PeriodicalIF":4.0,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144116618","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}

{"title":"5-Aminolevulinic Acid: from pyrrole biosynthetic precursor to multifunctional plant growth regulator","authors":"Liangju Wang,&nbsp;Jianting Zhang,&nbsp;Qingze Zhao,&nbsp;Liuzi Zhang","doi":"10.1016/j.jplph.2025.154524","DOIUrl":"10.1016/j.jplph.2025.154524","url":null,"abstract":"<div><div>5-Aminolevulinic acid (ALA) is a non-protein δ-amino acid and an essential precursor of tetrapyrrole compound biosynthesis. Nowadays, it is a well-known natural plant growth regulator with multiple biological regulatory functions. In this review, we summarize the regulatory effects of ALA in promoting plant growth and the development of organs such as roots, stems, leaves, flowers, and fruits under normal conditions as well as stressful conditions. We emphasize the newly revealed signaling transduction and transcriptional regulatory mechanisms of ALA in maintaining root functions against abiotic stresses, improving leaf photosynthetic performance, and enhancing fruit appearance and flavor qualities as well as storage. Although most of the current reports on ALA are still apparent effect descriptions rather than mechanism explorations, studies suggest that ALA can facilitate agricultural development toward higher yield, quality, efficiency, and safety. The regulatory mechanisms of ALA at different levels need further study in the future.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"310 ","pages":"Article 154524"},"PeriodicalIF":4.0,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144088696","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}

{"title":"Unveiling regional and altitudinal lipidomic analyte signatures of the argan tree (Argania spinosa L.) for environmental adaptation","authors":"El Faqer Abdelmoiz ,&nbsp;Rabeh Karim ,&nbsp;Rachidi Farid ,&nbsp;Assemar Fatima ezzahra ,&nbsp;Aasfar Abderrahim ,&nbsp;Filali-Maltouf Abdelkarim ,&nbsp;Belkadi Bouchra","doi":"10.1016/j.jplph.2025.154523","DOIUrl":"10.1016/j.jplph.2025.154523","url":null,"abstract":"<div><div>Environmental factors such as altitude, precipitation, and temperature shape the lipidomic profiles of the argan tree (<em>Argania spinosa</em> L.), supporting its adaptation to stress. This study investigated lipidomic profiling and pathways in argan tree leaves from four altitudinal zones (A: low, B: moderate, C: high, D: very high) across three Moroccan regions (Chtouka Aït Baha, Essaouira, and Tiznit) using Gas Chromatography-Mass Spectrometry (GC-MS). The GC-MS workflow included a transmethylation step that cleaves ester bonds and acetylations, yielding analytes derived from diverse precursor lipids such as glycerolipids, sterol esters, and wax esters. We identified 139 lipid analytes, categorized into fatty acyls (53 %), prenol lipids (41 %), and steroids (6 %). Shared lipids across all zones highlight core metabolic pathways essential for resilience, while unique lipids reflect zone-specific adaptations. Fourteen known analytes were identified as critical markers for regional adaptations through multivariate analyses, including Principal Component Analysis (PCA), Partial Least Squares Discriminant Analysis (PLS-DA), and Variable Importance in Projection (VIP) scores. Among these, three analytes (methyl 18-methyleicosanoate, Z,Z-11,13-Hexadecadien-1-ol, and 11-Octadecenoic acid) showed the highest accumulation in Zone A, whereas eleven analytes (Henicosyl formate, Dodecyl 2-methylbutanoate, Methyl 21-methyl-hexacosanoate, Methyl 13-methyltetradecanoate, Cetoleic acid, (Urs-12-en-3-ol, acetate, (3.beta.)-), Medicagenic acid, 2-(4a,8-Dimethyl-6-oxo-1,2,3,4,4a,5,6,8a-octahydro-naphthalen-2-yl)-propionaldehyde, A′-Neogammacer-22(29)-en-3-one, Pregna-5,17(20)-dien-3-ol, (3.beta.,17E)-, and estra-1,3,5(10)-trien-17-one, 3,4-bis(acetyloxy)- exhibited significant increases in Zone D. Multiple Linear Regression analysis showed that precipitation positively influenced analyte concentration (p = 0.00033), while altitude had a significant negative effect (p = 0.039). Pathways analysis highlighted the roles of cutin, suberin, and wax biosynthesis, as well as linoleic acid metabolism, in altitude-driven adaptations. This study demonstrates the metabolic plasticity of <em>Argania spinosa</em> L., offering insights for its conservation amidst climate change.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"311 ","pages":"Article 154523"},"PeriodicalIF":4.0,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144166825","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}

{"title":"Next generation technologies for protein structure determination: challenges and breakthroughs in plant biology applications","authors":"Veronica G. Maurino","doi":"10.1016/j.jplph.2025.154522","DOIUrl":"10.1016/j.jplph.2025.154522","url":null,"abstract":"<div><div>Advancements in structural biology have significantly deepened our understanding of plant proteins, which are central to critical biological functions such as photosynthesis, metabolism, signal transduction, and structural architechture. Gaining insights into their structures is crucial for unraveling their functions and mechanisms, which in turn has profound implications for agriculture, biotechnology, and environmental sustainability. Traditional methods in protein structural biology often fall short in addressing large protein assemblies and membrane proteins, and, in particular the dynamics and structural features of proteins in the native cellular context. This paper explores how next-generation technologies are transforming the field of plant protein structural biology, offering powerful tools to overcome longstanding obstacles and enabling remarkable scientific breakthroughs. Key technologies discussed include advanced X-ray crystallography, Cryo-Electron microscopy, Nuclear Magnetic Resonance spectroscopy, Cross-linking mass spectrometry, and Artificial Intelligence-driven approaches. These technologies are examined in terms of their challenges, innovations, and application with particular emphasis on their relevance to plant systems. Future directions in plant protein structural biology are also discussed. Although technical details are not covered in depth, readers are referred to the primary literature for more comprehensive information.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"310 ","pages":"Article 154522"},"PeriodicalIF":4.0,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144071960","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}

{"title":"Establishment of Agrobacterium-mediated genetic transformation and CRISPR/Cas9-guided gene editing in Elymus nutans","authors":"Cheng Li ,&nbsp;Xiaomei Peng ,&nbsp;Zhengshe Zhang ,&nbsp;Yaling Liu ,&nbsp;García-Caparrós Pedro ,&nbsp;Chunxiang Fu ,&nbsp;Yongping Yang ,&nbsp;Quanmin Dong ,&nbsp;Yuanwen Duan ,&nbsp;Xudong Sun","doi":"10.1016/j.jplph.2025.154513","DOIUrl":"10.1016/j.jplph.2025.154513","url":null,"abstract":"<div><div><em>Elymus nutans</em>, an allohexaploid (2n = 6x = 42) species with a StStHHYY genome, is a native perennial in the alpine grasslands of the Qinghai-Xizang Plateau, and has been widely used for artificial pasture and ecological restoration as a forage grass with highest yield on the plateau. Nevertheless, the lack of a stable transformation system has impeded further efforts to trait improvement of <em>E. nutans</em>. In the present study, we established a reliable <em>Agrobacterium</em>-mediated genetic transformation system for <em>E. nutans</em>, and successfully generated <em>EnTCP4</em>-edited plants using the CRISPR/Cas9 system. The editing efficiency achieved 19.23 % in <em>E. nutans</em>. Knocking out <em>EnTCP4</em> significantly delayed flowering and enhanced water-deficit stress resistance. This research represents a significant breakthrough in the genetic transformation and gene editing of <em>E. nutans</em>, laying a technological foundation to gain insight into gene functions and molecular breeding in <em>E. nutans</em>.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"310 ","pages":"Article 154513"},"PeriodicalIF":4.0,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143923846","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}

{"title":"Systematical accumulating and regulating evaluations of leaf functional metabolites in geographically isolated edible medicinal plants of Piper sarmentosum","authors":"Liu Ziting ,&nbsp;Wu Xiaoqing ,&nbsp;Wang Kemei ,&nbsp;Bai Yachao ,&nbsp;Guo Aimin ,&nbsp;Linan Huang ,&nbsp;Liao Bin ,&nbsp;Zhang Jun","doi":"10.1016/j.jplph.2025.154512","DOIUrl":"10.1016/j.jplph.2025.154512","url":null,"abstract":"<div><div>The edible medicinal plant <em>Piper sarmentosum</em> is widely distributed in south China. This study raised a hypothesis of geographically isolated <em>P</em>. <em>sarmentosum</em> plants possessing potential site- or/and plant-dependent accumulating metabolites, expressing genes, and colonizing bacteria. Here, <em>P</em>. <em>sarmentosum</em> plants of Guangzhou City (PG, comparison group) and Hainan Island (PH, control group) were collected for assaying leaf metabolomes (LMs), leaf transcriptomes (LTs), and leaf-assembled bacterial communities (LABCs), respectively. In LMs and LTs, 930 metabolites and 82,606 unigenes were identified with 552 differently accumulated metabolites (DAMs) and 28,177 differently expressed genes (DEGs), respectively. In LABCs, cluster analysis yielded 822 PG-PH-common, 1114 PG-unique, and 203 PH-unique operational taxonomic units (OTUs). In contrast of PH-LMs, the elevated accumulations of alkaloids and lipids and the decreased accumulations of flavonoids and phenolic acids were observed in PG-LMs. Typically, the DAMs and DEGs were co-enriched in two metabolic pathways of phenylpropanoids and flavonoids, visibly displaying the related DEGs, such as chalcone synthase (<em>CHS</em>), chalcone isomerase (<em>CHI</em>) and phenylalanine amino lyase (<em>PAL</em>), with regulating the functional DAMs, such as phenylalanine, tyrosine, p-coumaric acid, and naringenin. Noticeably, these DAMs were also significantly correlated with a number of different types or/and abundances of leaf-assembled bacteria (DTAB) between PG- and PH-LABCs, such as <em>Flavobacterium</em> and <em>Pseudomonas</em>. Therefore, this study clearly elucidated the functional metabolite accumulations and the close relationships with plant mRNA expressions and bacterial colonizations in geographically isolated plants of <em>P. sarmentosum</em>, providing new insight of selectively utilizing leaf food- and medicine-associated metabolites in different habitats of edible medicinal plants.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"310 ","pages":"Article 154512"},"PeriodicalIF":4.0,"publicationDate":"2025-05-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143932248","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}

{"title":"Drought-induced changes in antioxidant capacity, mineral content, and plant development in basal leafy cactus: comparisons between Pereskia aculeata miller and Pereskia bahiensis Gürke","authors":"Daniel S. Gomes ,&nbsp;Antônio N. Andrade ,&nbsp;Estephanni F.O. Dantas ,&nbsp;Vanessa A. Soares ,&nbsp;Damiana J. Araujo ,&nbsp;Sabrina K. Santos ,&nbsp;Adriano S. Lopes ,&nbsp;José E.S. Ribeiro ,&nbsp;Valquiria C.S. Ferreira ,&nbsp;Juliane M. Henschel ,&nbsp;Walter E. Pereira ,&nbsp;Thiago J. Dias ,&nbsp;Diego S. Batista","doi":"10.1016/j.jplph.2025.154503","DOIUrl":"10.1016/j.jplph.2025.154503","url":null,"abstract":"<div><div><em>Pereskia aculeata</em> is known for its high levels of proteins, minerals, vitamins, and bioactive compounds, qualifying it as an unconventional food. In contrast, the nutritional potential of <em>P. bahiensis</em> remains much less explored. Drought and defense-related bioregulators like salicylic acid (SA) and sodium nitroprusside (SNP) can influence plant composition and antioxidant responses, but their effects on <em>Pereskia</em> species are still unclear. This study aimed to assess the growth and nutritional profiles of <em>P. aculeata</em> and <em>P. bahiensis</em> under water deficit, SA, and SNP, testing whether <em>P. bahiensis</em> also holds food potential and whether these treatments enhance antioxidant capacity. For this, plants were grown under two irrigation regimes: well-watered (80 % water retention capacity – WRC) and water deficit (15 % WRC for 8 days, followed by total water restriction for 16 days). Additionally, plants were foliar sprayed with 100 μM SA, 100 μM SNP, or water (control). <em>P. bahiensis</em> exhibited greater phenolic compounds content and antioxidant capacity, but lower biomass production, and contents of sugars and minerals compared to <em>P. aculeata</em>. Drought increased the antioxidant capacity but decreased mineral content and biomass in the two <em>species;</em> while SA and SNP only affected Mn contents, not affecting the antioxidant capacity and growth. These findings highlight the great antioxidant capacity of <em>P. bahiensis</em>, surpassing that of <em>P. aculeata</em>, which underscores its potential as a nutraceutical plant. Moreover, water deficit can be used as a strategy to improve antioxidant capacity of both species, while SA and SNP treatments have no effects in their quality.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"309 ","pages":"Article 154503"},"PeriodicalIF":4.0,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143900088","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}

{"title":"The look insight – magnetic resonance imaging (MRI) of the inner life of plants","authors":"Ljudmilla Borisjuk ,&nbsp;Thomas Neuberger","doi":"10.1016/j.jplph.2025.154502","DOIUrl":"10.1016/j.jplph.2025.154502","url":null,"abstract":"<div><div>Magnetic resonance imaging (MRI) is one of the most versatile and widely used imaging techniques in modern medicine, but its incredible potential remains underutilized in plant science. Many aspects of the inner life of plants are still unknown and are waiting to be discovered by new innovative technological solutions. The ability of MRI to non-invasively explore processes inside living organisms offers an unparalleled opportunity to investigate metabolism, nutrient allocation, growth and development. Enabling the visualization of complex dynamics in living organisms in unprecedented ways could transform how we study and perceive plants. This article highlights the critical advancements and strategies that are paving the way for MRI to become an essential tool in plant research, unlocking new frontiers in biology and agriculture.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"309 ","pages":"Article 154502"},"PeriodicalIF":4.0,"publicationDate":"2025-04-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143900089","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}

{"title":"Selection and validation of reference genes for quantitative real-time PCR analysis across tissues at different developmental stages in Taraxacum kok-saghyz","authors":"Yongmei Li ,&nbsp;Yuan Yao ,&nbsp;Tiancheng Xu ,&nbsp;Xue Yang ,&nbsp;Yuting He ,&nbsp;Shengmin Zhang ,&nbsp;Qingbiao Xie ,&nbsp;Jie Cao ,&nbsp;Chaorong Tang ,&nbsp;Hui Liu","doi":"10.1016/j.jplph.2025.154501","DOIUrl":"10.1016/j.jplph.2025.154501","url":null,"abstract":"<div><div>Quantitative real-time polymerase chain reaction (qRT-PCR) is a highly sensitive and widely used method for analyzing gene expression profiles. Accurate qRT-PCR normalization requires the identification of stable reference genes under specific experimental conditions. Although seven reference genes have been used in <em>Taraxacum kok-saghyz</em> (TKS), an alternative natural rubber-producing crop, a systematic identification of reliable internal references for gene expression analysis across tissues at distinct developmental stages of TKS has not been conducted. In this study, we screened 12 candidate reference genes (CRGs) based on RNA-seq data from 26 TKS samples, representing five tissue types and nine developmental stages. The expression levels of the 12 CRGs, along with 7 previously reported reference genes (RRGs), were quantified by qRT-PCR across various tissues and developmental stages. The expression stability of the 19 genes was further evaluated by four commonly used algorithms (geNorm, NormFinder, comparative delta Ct, and BestKeeper), and their results were integrated by RefFinder to generate a comprehensive stability ranking. The final results revealed that <em>TkADF1</em> and <em>TkRPT6A</em> were the most suitable internal control genes for the all-tissue group and leaf samples. <em>TkUPL</em> and <em>TkSIZ1</em> were found to be optimal for root samples, while <em>TkADF1</em> and <em>TkSRPRA</em> were preferred choices for latex samples. Moreover, validation using two rubber biosynthesis-related genes (<em>TkFPS1</em> and <em>TkSRPP2</em>) confirmed the reliability of these recommended genes, showing a strong positive correlation with the RNA-seq data. This study provides reliable reference genes for qRT-PCR normalization in TKS, facilitating future research on developmental regulation and natural rubber biosynthesis.</div></div>","PeriodicalId":16808,"journal":{"name":"Journal of plant physiology","volume":"309 ","pages":"Article 154501"},"PeriodicalIF":4.0,"publicationDate":"2025-04-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143903919","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}