中国农业科学 ›› 2025, Vol. 58 ›› Issue (7): 1333-1343.doi: 10.3864/j.issn.0578-1752.2025.07.007

• 植物保护 • 上一篇    下一篇

广泛非靶向代谢组学解析小麦抗条锈病反应中叶绿体代谢产物

吴郁(), 曲翔汝, 杨丹, 伍芩, 陈国跃, 江千涛, 魏育明(), 许强()   

  1. 四川农业大学小麦研究所/西南作物基因探索与利用国家重点实验室,成都 611130
  • 收稿日期:2024-12-06 接受日期:2025-02-22 出版日期:2025-04-08 发布日期:2025-04-08
  • 通信作者:
    魏育明,E-mail:
    许强,E-mail:
  • 联系方式: 吴郁,E-mail:1363797694@qq.com。
  • 基金资助:
    国家自然科学基金青年科学基金项目(32102229)

Widespread Non-Targeted Metabolomics Reveals Metabolites of Chloroplasts in Wheat Responses to Stripe Rust

WU Yu(), QU XiangRu, YANG Dan, WU Qin, CHEN GuoYue, JIANG QianTao, WEI YuMing(), XU Qiang()   

  1. Triticeae Research Institute/State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China, Sichuan Agricultural University, Chengdu 611130
  • Received:2024-12-06 Accepted:2025-02-22 Published:2025-04-08 Online:2025-04-08

摘要:

【目的】揭示小麦抗条锈病反应中叶绿体代谢物变化,明确小麦叶绿体关键代谢产物在抗条锈病中的作用。【方法】以小麦品种水源11为试验材料,设置喷施无菌水(对照组)和接种条锈菌(Puccinia striiformis f. sp. tritici)生理小种CYR23(试验组),提取喷施后48和72 h(hour post infection,hpi)的小麦叶绿体,利用超高液相色谱串联质谱(UHPLC-MS/MS)进行广泛非靶向代谢组学分析,通过主成分分析和相关性分析展示各组间和组内样本的差异性与相关性,并采用正交偏最小二乘判别分析(OPLS-DA)模型的VIP(variable importance in projection)值来评估抗病组与对照组的叶绿体代谢物差异;通过京都基因与基因组百科全书(KEGG)数据库对叶绿体差异代谢物进行通路富集分析,探究小麦在条锈病抗性反应中叶绿体的主要代谢通路。【结果】代谢组数据质量可靠,组内样品重复性好,且组间样品存在差异。共检测到1 496种代谢物,主要包括脂质及类脂分子、有机酸及其衍生物、有机杂环化合物、苯丙类和聚酮类物质、有机氧化物、苯类化合物等。不同组间代谢物种类较为相似,但在不同侵染时间代谢物含量变化显著。在48 hpi与对照组(CK)中,差异代谢物有121种,包括21种上调代谢物和100种下调代谢物;72 hpi与CK组中,差异代谢物有58种,其中35种上调,23种下调;72与48 hpi组中,差异代谢物有53种,其中33种上调,20种下调。不同侵染时间48 hpi vs CK与72 hpi vs CK也共有4种差异代谢物。KEGG分析发现差异代谢物富集到多条代谢通路,其中亚油酸代谢通路中的亚油酸在接种条锈菌48和72 h后均上调表达,两个时间点分别上调表达2.75和2.93倍,表明亚油酸受条锈菌诱导持续合成。【结论】叶绿体中响应小麦抗条锈病的主要代谢物有脂质和脂肪酸及其衍生物、有机酸及其衍生物、有机氧化物和苯丙类、聚酮类物质,其中亚油酸可能在小麦持续抵抗条锈病过程中起重要作用。

关键词: 小麦, 条锈病, 条形柄锈菌, 叶绿体, 代谢组学, 亚油酸

Abstract:

【Objective】The purpose of this study is to elucidate the changes of chloroplast metabolites during wheat resistance to stripe rust, and to clarify the role of key chloroplast metabolites in wheat resistance to stripe rust.【Method】Wheat cultivar Suwon11 was used as the experimental material, wheat chloroplasts were extracted 48 and 72 h after spraying sterile water (control group) and Puccinia striiformis f. sp. tritici CYR23 (experimental group), and a widespread non-targeted metabolomics analysis was conducted via ultra-high performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Principal component analysis (PCA) and correlation analysis were employed to visualize inter-group and intra-group differences and associations. VIP (variable importance in projection) values derived from the orthogonal partial least squares discriminant analysis (OPLS-DA) model were employed to identify differential chloroplast metabolites between the disease-resistant and control groups. Pathway enrichment analysis was performed on differential chloroplast metabolites using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database to investigate key metabolic pathways in chloroplasts during the resistance response of wheat to stripe rust.【Result】The metabolomics data exhibited high quality, with good intra-group reproducibility and significant inter-groups variability validated by multivariate statistical analyses. A total of 1 496 metabolites were detected, which predominantly included lipids and lipid-like molecules, organic acids and their derivatives, organic heterocyclic compounds, phenylpropanoids and polyphenolic compounds, organic oxides, and benzoid compounds. The metabolite species were generally similar between different groups, but significant variations in metabolite levels were observed at different infection time points. At 48 hours post infection (hpi) compared to the control group (CK), 121 differential metabolites were identified, including 21 up-regulated and 100 down-regulated metabolites; At 72 hpi compared to the CK, 58 differential metabolites were detected, with 35 up-regulated and 23 down-regulated; Between 72 and 48 hpi groups, 53 differential metabolites were observed, of which 33 were up-regulated and 20 were down-regulated. Four differential metabolites were shared between the 48 hpi vs CK and 72 hpi vs CK comparisons. KEGG analysis revealed that the differential metabolites were enriched in multiple metabolic pathways. Among them, the expression of linoleic acid which in the linoleic acid metabolic pathway exhibited 2.75- and 2.93- fold increases after inoculation 48 and 72 h, respectively, indicating that linoleic acid was continuously synthesized induced by P. striiformis f. sp. tritici.【Conclusion】The chloroplast metabolites associated with wheat resistance to stripe rust primarily include lipids, fatty acids and their derivatives, organic acids and their derivatives, organic oxides, as well as phenylpropanoids and polyphenolic compounds. Notably, linoleic acid may play a crucial role in the sustained resistance of wheat against stripe rust.

Key words: wheat (Triticum aestivum), stripe rust, Puccinia striiformis f. sp. tritici, chloroplast, metabonomics, linoleic acid