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Journal of Integrative Agriculture
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Characterization of defense responses in forage and food barley hosts during interaction with Epichloë endophyte

Kamran Malik*, Fangli Wei*, Taixiang Chen#, Chunjie Li#

 State Key Laboratory of Herbage Improvement and Grassland Agro-ecosystems, Key Laboratory of Grassland Livestock Industry Innovation, Ministry of Agriculture and Rural Affairs, Engineering Research Center of Grassland Industry, Ministry of Education; Gansu Tech Innovation Centre of Western China Grassland Industry; Centre for Grassland Microbiome, College of Pastoral Agriculture Science and Technology, Lanzhou University, Lanzhou 730000, China 

 Highlights 

§ Endophyte colonization was significantly higher in wild barley (54.21%) than in cultivated barley (9.91%).

§ Transcriptomics profiling revealed host metabolic reprogramming against endophyte stress.

§ Endophyte triggered a pathogen-like defense response in cultivated host, which mounted an earlier and stronger defense with elevated MAPK/ROS.

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摘要  

内生菌普遍存在于植物中,对植物的生长发育起着重要作用。本研究将禾草内生真菌Epichloë bromicola菌株WBE1人工接种到野生大麦(天然宿主Hordeum brevisubulatum)和栽培大麦(非天然宿主Hordeum vulgare)体内,构建了内生真-野大麦共生体和内生真-大麦共生体,测定了天然宿主和非天然宿主的胼胝质、木质素、细胞死亡率、早期信号分子、第二信使内源信号分子等生理性状对内生真菌WBE1定殖后的响应,以及不同定殖时期差异基因的表达模式。研究结果表明,E. bromicola菌株WBE1在野生大麦和栽培大麦体内的定殖成功率分别为 54.21%和9.91%。人工接种内生真菌刺激了两种宿主植物的胼胝质含量、木质素含量的累积和细胞死亡率,但栽培大麦的反应较野生大麦更强烈。内生真菌的侵染诱导了早期信号分子的表达,诱导了两种宿主的O2- 产生速率以及 H2O2和NO的累积。在侵染早期,栽培大麦的丝裂原活化蛋白激酶(MAPK)活性在侵染后第2天和第4天分别比野生大麦提12.24%和54.60%。转录组学分析显示,与野生大麦相比,栽培大麦触发了更早、更有针对性的防御反应,涉及抗性相关次生代谢物生物合成和关键信号分子的基因在不同阶段特异性上调,而与氧脂素生物合成、脂质氧化、细胞对环境刺激的反应、氧化还原酶活性和血红素结合相关的基因下调表达。本研究结果表明,禾草内生真菌E. bromicola侵染能有效地诱导栽培大麦增强早期防御反应,与天然宿主野生大麦相比,内生真菌对非天然宿主栽培大麦防御反应的诱导作用更强烈且更迅速。



Abstract  

Endophytes are prevalent in plants and significantly contribute to plant growth and development. In the present study, Epichloë bromicola endophyte strain WBE1 was artificially inoculated into wild barley (Hordeum brevisubulatum, natural host) and cultivated barley (Hordeum vulgare, novel host) to obtain endophyte-barley symbionts. Physiological traits, such as callus formation, lignin content, cell mortality, early signaling molecules, second messenger endogenous signaling molecules, and the expression patterns of differential genes at different time periods were studied. The colonization rate of E. bromicola was 54.21% in wild barley and 9.91% in cultivated barley. Artificial endophytic infection enhanced callus growth, lignin content, and cell mortality in both hosts, with cultivated barley showing stronger resistance than wild barley. The infection induced the expression of early signaling molecules, and the O2- production rate as well as H2O2 and NO contents were increased in both hosts. During the early infection stage, mitogen-activated protein kinase (MAPK) activity in cultivated barley increased by 12.24% and 54.60% compared with wild barley at 2 and 4 days post-infection, respectively. Transcriptomic analysis revealed that cultivated barley triggered an earlier and targeted defense response than wild barley, characterized by the stage-specific upregulation of genes involved in resistance-related secondary metabolite biosynthesis and key signaling molecules. Expression patterns showed upregulation of signaling molecules alongside downregulation of genes associated to oxylipin biosynthesis, lipid oxidation, cellular responses to environmental stimuli, oxidoreductase activity, and heme binding. These findings indicated that E. bromicola infection effectively triggered an enhanced and timely defense response in cultivated barley. 

Keywords:  fungal endophyte       inoculation       signaling molecules              secondary metabolites              gene expression  
Online: 10 February 2026  
Fund: 

This research was funded by the National Science Foundation of China (U21A20239 and 32471779); the Science and Technology Planning Project of Gansu Province, China (Natural Science Foundation) (22JR5RA458).

About author:  Kamran Malik, E-mail: malik@lzu.edu.cn; Fangli Wei, E-mail: 1937527612@qq.com; #Correspondence Chunjie Li, E-mail: chunjie@lzu.edu.cn; Taixiang Chen, E-mail: chentx@lzu.edu.cn * These authors contributed equally to this study.

Cite this article: 

Kamran Malik, Fangli Wei, Taixiang Chen, Chunjie Li. 2026. Characterization of defense responses in forage and food barley hosts during interaction with Epichloë endophyte. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2026.02.023

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